Metabolic engineering of indole glucosinolates in Chinese cabbage hairy roots expressing Arabidopsis CYP79B2, CYP79B3, and CYP83B1 |
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Authors: | Yun-Xiang Zang Doo-Hwan Kim Beom-Seok Park Seung-Beom Hong |
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Institution: | (1) Faculty of Life Science, University of Copenhagen, 1871 Frederiksberg C, Denmark;(2) Boyce Thompson Institute for Plant Research, 1 Tower Road, Ithaca, NY 14853, USA;(3) Present address: Monsanto Company, St. Louis, MO, USA; |
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Abstract: | Indole glucosinolates (IG), a group of secondary metabolites found almost extensively in the order Brassicales, play an important
role in the interaction between plant and insect or microorganism. In order to explore the possibility of IG metabolic engineering
in Chinese cabbage hairy roots, three Arabidopsis cDNAs CYP79B2, CYP79B3, and CYP83B1 combined with rolABC were introduced into Chinese cabbage by Agrobacterium-mediated transformation. Overexpression of CYP79B2, CYP79B3, or CYP83B1 alone did not affect the accumulation levels of IG
in transgenic hairy roots. However, when CYP83B1 was overexpressed together with CYP79B2 and/or CYP79B3, some of the transgenic
hairy roots accumulated higher levels of glucobrassicin (GBC) or 4-methoxy glucobrassicin (4-OMeGBC) than control hairy root
line carrying rolABC vector. With regard to the IG accumulation, overexpression of all three cDNAs showed no better than overexpression of both
cDNAs. Both 4-OMeGBC and neoglucobrassicin (neo-GBC) were found to be the main components of IG that comprise about 90% of
total IG in all types of Chinese cabbage hairy root lines. In transgenic hairy root lines rB3B1-8 and rB2B1B3-5, 4-OMeGBC
increased to 2 and 1.5-fold, while neo-GBC decreased to 0.5 and 0.6-fold, respectively. This suggests that an increased production
of 4-OMeGBC causes a reduction of neo-GBC level since the two types derive from a common precursor GBC. However, in terms
of the total IG level, the transgenic hairy roots did not show significant differences from controls. |
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